Biological implications of 2-chlorocyclohexa-2,5-diene-1,4-dione toward ribonuclease A

Albert R. Vaughn, Caitlin B. Redman, Sophia M. Kang, Jisook Kim

Department of Chemistry, University of Tennessee at Chattanooga, Chattanooga, USA.

DOI: 10.4236/abb.2013.41004                

Abstract

2-Chlorocyclohexa-2,5-diene-1,4-dione (CBQ) or 2-chloro1,4-benzquinone is one of the common metabolites of polycyclic aromatic hydrocarbons generated through industrial processes. This report describes the biological effects of CBQ toward ribonuclease A (RNase). We also investigated the inhibition of RNase modifications and the reactivity of CBQ toward selected amino acids. The study was carried out by incubating RNase or amino acids with CBQ in a concentration- and a time-dependent manner at 37°C and pH 7.0. SDS-PAGE results showed oligomerization as well as polymeric aggregation of RNase when incubated with CBQ as early as in 10 min. CBQ-induced RNase modifications were inhibited in the presence of NADH or ascorbic acid. CBQ reactivity toward selected amino acids was also evaluated by determining the second-order rate constants for the reactions of CBQ with selected amino acids. It was found that the reactivity toward CBQ decreased in the order of lysine > threonine > serine >> aspartate > cysteine.

Keywords

Chlorobenzoquinone; PAH; Protein Modification; Ribonuclease A

Conflicts of Interest

The authors declare no conflicts of interest.

References

[1]	Hrudey, S.E., (2009) Chlorination disinfection by-products, public health risk tradeoffs and me. Water Research, 43, 2057-2092. doi:10.1016/j.watres.2009.02.011
[2]	Morbt, N., Tomm, J., Feltens, R., Mogel, I., Kalkhof, S., Murugesan, K., Wirth, H., Vogt, C., Binder, H., Lehmann, I. and von Bergen, M. (2010) Chlorinated benzenes cause concomitantly oxidative stress and induction of apoptotic markers in lung epithelial cells (A549) at nonacute toxic concentrations. Journal of Proteome Research, 10, 363-378. doi:10.1021/pr1005718
[3]	Ogata, M., Taguchi, T., Hirota, N., Shimada, Y. and Nakae, S. (1991) Quantitation of urinary chlorobenzene metabolites by HPLC: Concentrations of 4-chlorocatechol and chlorophenols in urine and of chlorobenzene in biological specimens of subjects exposed to chlorobenzene. International Archives of Occupational and Environmental Health, 63, 121-128. doi:10.1007/BF00379075
[4]	Nair, R.S., Barter, J.A., Schroeder, R.E., Knezevich, A. and Stack, C.R. (1987) A two-generation reproduction study with monochlorobenzene vapor in rats. Fundamental and Applied Toxicology, 9, 678-686. doi:10.1016/0272-0590(87)90174-6
[5]	Aiso, S., Takeuchi, T., Arito, H., Nagano, K., Yamamoto, S. and Matsushima, T. (2005) Carcinogenicity and chronic toxicity in mice and rats exposed by inhalation to paradichlorobenzene for two years. Journal of Veterinary Medical Science, 67, 1019-1029. doi:10.1292/jvms.67.1019
[6]	Yamazaki, K., Aiso, S., Matsumoto, M., Kano, H., Arito, H., Nagano, K., Yamamoto, S. and Matsushima, T. (2006) Carcinogenicity and chronic toxicity of 1,4-dichloro-2-nitrobenzene in rats and mice by two years feeding. Industrial Health, 44, 230-243. doi:10.2486/indhealth.44.230
[7]	Sharma, S., Mukhopadhyay, M. and Murthy, Z.V.P. (2010) Degradation of 4-chlorophenol in wastewater by organic oxidants. Industrial & Engineering Chemistry Research, 49, 3094-3098. doi:10.1021/ie9018066
[8]	Zhao, Y., Qin, F., Boyd, J.M., Anichina, J. and Li, X.F. (2010) Characterization and determination of chloro- and bromo-benzoquinones as new chlorination disinfection byproducts in drinking water. Analytical Chemistry, 82, 4599-4605. doi:10.1021/ac100708u
[9]	Qin, F., Zhao, Y.Y., Zhao, Y., Boyd, J.M., Zhou, W. and Li, X.F. (2010) A toxic disinfection by-product, 2,6-di-chloro-1,4-benzoquinone, identified in drinking water. Angewandte Chemie International Edition, 49, 790-792. doi:10.1002/anie.200904934
[10]	Yin, L., Niu, J., Shen, Z. and Chen, J. (2010) Mechanism of reductive decomposition of pentachlorophenol by Ti-doped beta-Bi2O3 under visible light irradiation. Environmental Science & Technology, 44, 5581-5586. doi:10.1021/es101006s
[11]	Yin, L., Shen, Z., Niu, J., Chen, J. and Duan, Y. (2010) Degradation of pentachlorophenol and 2,4-dichlorophenol by sequential visible-light driven photocatalysis and laccase catalysis. Environmental Science & Technology, 44, 9117-9122. doi:10.1021/es1025432
[12]	Snyder, R. and Hedli, C.C. (1996) An Overview of Benzene Metabolism. Environmental Health Perspectives, 104, 1165-1172.
[13]	Bolton, J.L., Trush, M.A., Penning, T.M., Dryhurst, G. and Monks, T.J. (2000) Role of quinones in toxicology. Chemical Research in Toxicology, 13, 135-160. doi:10.1021/tx9902082
[14]	Kondrova, E., Stopka, P. and Soucek, P. (2007) Cytochrome P450 destruction by benzene metabolites 1,4-benzoquinone and 1,4-hydroquinone and the formation of hydroxyl radicals in minipig liver microsomes. Toxicology in Vitro, 21, 566-575. doi:10.1016/j.tiv.2006.11.002
[15]	Bodell, W.J., Pathak, D.N., Levay, G., Ye, Q. and Pongracz, K. (1996) Investigation of the DNA adducts formed in B6C3F1 mice treated with benzene: Implications for molecular dosimetry. Environmental Health Perspectives, 104, 1189-1193.
[16]	Buben, A., Narasimhan, N. and Hanzlik, R.P. (1988) Effects of chemical and enzymic probes on microsomal covalent binding of bromobenzene and derivatives. Evidence for quinones as reactive metabolites. Xenobiotica, 18, 501-510. doi:10.3109/00498258809041687
[17]	Anichina, J., Zhao, Y., Hrudey, S.E., Le, X.C. and Li, X.F. (2010) Electrospray ionization mass spectrometry characterization of interactions of newly identified water disinfection byproducts halobenzoquinones with oligodeoxynucleotides. Environmental Science & Technology, 44, 9557-9563. doi:10.1021/es1024492
[18]	Nguyen, T.N., Bertagnolli, A.D., Villalta, P.W., Buhlmann, P. and Sturla, S.J. (2005) Characterization of a deoxyguanosine adduct of tetrachlorobenzoquinone: Dichloro-benzoquinone-1,N2-etheno-2’-deoxyguanosine. Chemical Research in Toxicology, 18, 1770-1776. doi:10.1021/tx050204z
[19]	Slaughter, D.E. and Hanzlik, R.P. (1991) Identification of epoxide- and quinone-derived bromobenzene adducts to protein sulfur nucleophiles. Chemical Research in Toxicology, 4, 349-359. doi:10.1021/tx00021a015
[20]	Hanzlik, R.P. (1986) Chemistry of covalent binding: Studies with bromobenzene and thiobenzamide. Advances in Experimental Medicine and Biology, 197, 31-40. doi:10.1007/978-1-4684-5134-4_3
[21]	Zaborska, W., Krajewska, B., Kot, M. and Karcz, W. (2007) Quinone-induced inhibition of urease: Elucidation of its mechanisms by probing thiol groups of the enzyme. Bioorganic Chemistry, 35, 233-242. doi:10.1016/j.bioorg.2006.11.001
[22]	Meade, S.J., Miller, A.G. and Gerrard, J.A. (2003) The role of dicarbonyl compounds in non-enzymatic crosslinking: A structure-activity study. Bioorganic & Medicinal Chemistry, 11, 853-862. doi:10.1016/S0968-0896(02)00564-3
[23]	Kim, J., Vaughn, A.R., Cho, C., Albu, T.V. and Carver, E.A. (2012) Modifications of ribonuclease A induced by p-benzoquinone. Bioorganic Chemistry, 40, 92-98. doi:10.1016/j.bioorg.2011.11.002
[24]	Reznick, A.Z. and Packer, L. (1994) Oxidative damage to proteins: Spectrophotometric method for carbonyl assay. Methods in Enzymology, 233, 357-363. doi:10.1016/S0076-6879(94)33041-7
[25]	Laemmli, U.K. (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature, 227, 680-685. doi:10.1038/227680a0
[26]	Weber, K. and Osborn, M. (1969) The reliability of molecular weight determinations by dodecyl sulfate polyacrylamide gel electrophoresis. The Journal of Biological Chemistry, 244, 4406-4412.
[27]	Yuasa, J., Yamada, S. and Fukuzumi, S. (2008) One-step versus stepwise mechanism in protonated amino acid-promoted electron-transfer reduction of a quinone by electron donors and two-electron reduction by a dihydronicotinamide adenine dinucleotide analogue. Interplay between electron transfer and hydrogen bonding. Journal of the American Chemical Society, 130, 5808-5820. doi:10.1021/ja8001452
[28]	Fukuzumi, S., Fujii, Y. and Suenobu, T. (2001) Metal ion-catalyzed cycloaddition vs hydride transfer reactions of NADH analogues with p-benzoquinones. Journal of the American Chemical Society, 123, 10191-10199. doi:10.1021/ja016370k
[29]	Gorner, H. (2005) Photoreactions of 1,4-Naphtho-quinones: Effects of substituents and water on the intermediates and reactivity. Photochemical & Photobiological Sciences, 81, 376-383. doi:10.1562/2004-08-11-RA-270.1
[30]	Verrax, J., Delvaux, M., Beghein, N., Taper, H., Gallez, B. and. Buc Calderon, P (2005) Enhancement of quinone redox cycling by ascorbate induces a caspase-3 independent cell death in human leukaemia cells. An in vitro comparative study. Free Radical Research, 39, 649-657. doi:10.1080/10715760500097906
[31]	Roginsky, V.A., Barsukova, T.K and Stegmann, H.B. (1999) Kinetics of redox interaction between substituted quinones and ascorbate under aerobic conditions. Chemico-Biological Interactions, 121, 177-197. doi:10.1016/S0009-2797(99)00099-X
[32]	Person, M.D., Mason, D.E., Liebler, D.C., Monks, T.J. and Lau, S.S. (2005) Alkylation of cytochrome c by (glutathion-S-yl)-1,4-benzoquinone and iodoacetamide demonstrates compound-dependent site specificity. Chemical Research in Toxicology, 18, 41-50. doi:10.1021/tx049873n
[33]	Hanzlik, R.P., Harriman, S.P. and Frauenhoff, M.M. (1994) Covalent binding of benzoquinone to reduced ribonuclease. Adduct structures and stoichiometry. Chemical Research in Toxicology, 7, 177-184. doi:10.1021/tx00038a010